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AAT Bioquest/ReadiUse™ mammalian cell lysis buffer *5X*/20012/10 mL
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | None/None |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | R |
| Category | MicroBIOLOGy FluorescenceImaging |
| Related | BiochemicalAssays |
1 WarmupReadiUse™mammaliancelllysisbuffer*5X*atroomtemperaturebeforeuse.
2 Prepare1XReadiUse™mammaliancelllysisbufferbyadding1mlof5XReadiUse™mammaliancelllysisbufferinto4mlofMili-Qwater,andmixthemwell.
3 The1XReadiUse™mammaliancelllysisbuffer(fromStep2)nowisreadytouse.
4 Treatcellsasdesired,washplatewithPBStoremoveresidualmedia.
5 Thefollowingtableisthesuggestedvolumeforlysisadherentcells.
Plate | 384-well | 96-well | 48-well | 24-well | 12-well | 6-well | 100cm2 |
Volume | 20µL | 50µL | 100µL | 150µL | 200mL | 300mL | 800mL |
6 Lysenon-adherentcellsbyaddingabout100µLof1XReadiUse™mammaliancelllysisbufferto1-5X106cellpellet.
7 Incubateatroomtemperaturefor10-20min.
8 Usethecelllysatedirectlyorcentrifugeitat1500rpmfor5minutes,usethesupernatantfor
9 furtherbiochemicalanalysis.
10 Thecelllysisbuffercanalsobeusedforlysebacterialorplanttissues.Seetheappendixfordetails.
Appendix:TestSamplePreparationsUsing1XCellLysisBuffer
1. PlantCellSamples:
Homogenizeleavewiththelysisbufferat200mg/mL,andcentrifugeat2500rpmfor5-10minutes,usethesupernatantfortests.
2. BacterialCellSamples:
Collectbacterialcellsbycentrifugation((10,000g,0°C,15min).Useabout 100to10millioncells/mLlysisbuffer,keepthetreatedsolutionatroomtemperaturefor15minutes. Centrifugeat2500rpmfor5minutes,andusethesupernatantfortests.
3. TissueSamples:
Weigh~20mgtissue,washwithcoldPBS.Homogenizewith400μloflysisbufferinamicro-centrifugetube.Centrifugeat2500rpmfor5-10minutes,usethesupernatantfortheassay.
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